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Related Experiment Videos

Cloning the gene for ribonuclease E, an RNA processing enzyme.

A Ray, D Apirion

    Gene
    |December 1, 1980
    PubMed
    Summary

    Researchers isolated a bacteriophage lambda Ch25rne+ that carries the gene for ribonuclease E (RNase E) in E. coli. This discovery aids in understanding bacterial RNA processing and RNase E function.

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    Area of Science:

    • Molecular Biology
    • Bacteriology
    • Genetics

    Background:

    • Ribonuclease E (RNase E) is crucial for bacterial RNA processing in E. coli.
    • Understanding RNase E function requires tools for its genetic manipulation and study.

    Purpose of the Study:

    • To isolate and characterize a transducing bacteriophage carrying the E. coli ribonuclease E gene.
    • To develop a tool for studying RNase E function and its role in RNA processing.

    Main Methods:

    • Construction of a lambda Charon 25 bacteriophage library with random Escherichia coli HindIII fragments.
    • Screening for lambda Ch25rne+ by its ability to complement a temperature-sensitive rne-3071 mutant.
    • Analysis of RNase E levels, polypeptide synthesis, and host DNA insert size.

    Main Results:

    • Isolation of lambda Ch25rne+, a transducing bacteriophage encoding E. coli RNase E.
    • Successful complementation of a temperature-sensitive rne-3071 mutant, restoring normal RNA processing at 45°C.
    • Demonstration of doubled RNase E levels in lysogenized strains and synthesis of a 71,000 m.wt. polypeptide.

    Conclusions:

    • Lambda Ch25rne+ is a functional tool for studying RNase E.
    • The isolated phage carries the gene for RNase E, approximately 1.9 kb in size.
    • This provides a new avenue for investigating bacterial RNA metabolism and RNase E function.

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